Capturing Spatial Heterogeneity Using UAV-Derived Multispectral Phenotypes and Random Regression Models Improves Genetic Evaluation in Alfalfa Breeding Trials

Abstract

Spatial heterogeneity is a major source of environmental noise in field trials, reducing genetic evaluation accuracy. Conventional spatial correction methods use field layout and plot proximity but often fail to capture in-season dynamics or leverage temporal data from high-throughput phenotyping. We hypothesized that longitudinal UAV-derived multispectral measurements could characterize field-scale environmental heterogeneity and improve genetic evaluation in breeding trials. Multispectral imagery was collected throughout the growing season across eight alfalfa trials in New York and New Mexico (20222023). Vegetation indices were evaluated for heritability and association with biomass yield using univariate and multi-trait models. Longitudinal measurements were modeled with random regression (RR) to estimate plot-specific permanent environmental effects (PEE), incorporated into genomic mixed models within a two-stage framework. Model performance was compared with conventional autoregressive (AR) and baseline (no correction) models using metrics of consistency and accuracy of estimated genetic effects across replicates. Vegetation indices showed moderate to high heritability, with GNDVI consistently outperforming NDVI and NDRE. Both univariate and multi-trait analyses revealed substantial spatial variability, particularly in New Mexico trials. RR-derived PEE captured field-scale spatial patterns and improved genetic signal recovery relative to baseline and AR models, with the largest gains under strong spatial heterogeneity and comparable performance where variability was limited. The RR framework improved reproducibility of genetic effect estimates across independent field subsets without compromising agreement with full-trial evaluations. These findings show that integrating UAV data via random regression enhances genetic signal recovery and selection accuracy, with broader potential for precision agriculture and biosensor-driven crop management.